Proteolytic enzymes as structural probes for ribonuclease BS-1.
Trypsin, pepsin and subtilisin have been used as conformational probes for the structure of bovine seminal ribonuclease BS-1 by studying, under definite conditions, their effects on the seminal enzyme, a dimeric protein made up to two identical subunits; on bovine pancreatic monomeric ribonuclease A (EC 3.1.4.22) with a polypeptide chain homologous to that of the seminal ribonuclease subunit chain; and on a monomeric, active and stable derivative of seminal ribonuclease. The results show: (1) that the C-terminal regions of the pancreatic and the seminal proteins are very similar as they appear to fit in an identical way to the active site of pepsin; (2) that the resistance of the N-terminal region of ribonuclease BS-1 to subtilisin is not due to the dimeric structure of the protein, but to the conformation of this region, where an essential feature is the presence of a proline residue at position 19; (3) that the monomer of ribonuclease BS-1 is resistant to tryptic action only when bound to the partner monomer in the quaternary structure of the protein. This indicates that dissociation of the seminal ribonuclease makes some potentially susceptible susceptible bond or bonds available to trypsin either through a conformational change of the protein subunit, or by simply exposing the protein area hidden at the intersubunit interfaces.